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Advances in Chemistry Applied to Forensic Science

Article  in  Chemical Society Reviews · March 2006

DOI: 10.1039/b415890n · Source: PubMed

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TUTORIAL REVIEW www.rsc.org/csr | Chemical Society Reviews

Advances in chemistry applied to forensic science

David F. Rendle
Received 1st August 2005
First published as an Advance Article on the web 12th October 2005
DOI: 10.1039/b415890n

Acts of terrorism, an increase in the use of firearms, drug abuse, the use of so-called date-rape
drugs, and driving whilst under the influence of drugs, are just some of the subjects frequently in
the news. In the absence of fingermarks and of material leading to the recovery of DNA, the
forensic scientist has to rely upon chemical analysis of trace amounts of materials including
explosives, drugs, toxicological specimens, firearms discharge residues, fibres, glass, paint, soil
etc., in order to establish or eliminate links between suspect and victim and/or scene. This tutorial
review describes analytical problems facing the forensic chemist, and the current methods and
techniques employed to tackle them.

Introduction Fingermarks at the scene of a crime are also individual-

Of late the general public has become more aware of the
application of science to crime solving, due mainly to the
proliferation of television programmes, both documentary and
fictional, and accounts of high-profile court cases in national
newspapers, each describing detailed aspects of forensic
science. Acts of terrorism, an increase in criminal use of
firearms, drug trafficking, the use of so-called date-rape drugs,
and driving whilst under the influence of drugs, are just some
of the subjects frequently in the news.
Since the discovery of individual-specific ‘‘fingerprints’’ of
human DNA and subsequent forensic application of DNA
fingerprints1 in 1985, forensic science laboratories have
committed considerable resources to the development of
DNA profiling methods. The highly-specific nature of DNA
evidence renders it invaluable as a means of identification.
specific, but however careful the criminal may be to avoid
leaving his fingerprints, inadvertent contact with other parts of
his person may result in his leaving body fluids or tissue at the
scene, each of which will yield his DNA. In the absence of
fingermarks and of material leading to recovery of DNA,
however, the forensic scientist has to rely upon chemical
analysis of trace amounts of materials including explosives,
drugs, firearms discharge residues, fibres, glass, paint, soil etc.,
in order to establish or eliminate links between suspect and
victim and/or scene. This review will examine the current
methods of forensic analytical chemistry for evidence types
including those listed above. General textbooks such as those
of White2 and Saferstein3 provide excellent introductory
information.
Firearms discharge residues

Scientific Consultant, 9 Wiltshire Drive, Wokingham, Berkshire,
UK RG40 1TQ. E-mail: d.rendle@virgin.net
David Rendle obtained his
PhD in Chemistry at the
University of Guelph,
Ontario, Canada in 1972.
After a Teaching Post-
doctoral Fellowship at UBC
Vancouver, he joined the
Metropolitan Police Forensic
Science Laboratory in London
in 1975, spending fifteen years
as Head of its X-ray Section
(Diffraction & Radiography)
and six years in its R&D
Section. He continued in
David Rendle R&D with the UK Forensic
Science Service from 1996
until 2003. He is currently a Senior Visiting Fellow in the
Department of Chemistry at the University of Reading, and is
also an External Examiner for the MSc course in Forensic
Engineering & Science at Cranfield University, RMCS,
Shrivenham.
The gases and particulate matter generated when a firearm is
used are known as firearms discharge residues (FDR). In order
to understand why and how they arise, it is necessary to
consider the make up of a typical cartridge. A cartridge case
contains three components: primer, propellant and projectile
(bullet). The primer is ignited by the percussive effect of the
weapon’s firing pin striking the primer cap on the end of the
cartridge. The primer burns rapidly and ignites the propellant,
which in turn burns very quickly, producing a large volume of
gases inside the cartridge case which forces the bullet down the
barrel. When a weapon is fired, in addition to the forward
propulsion of the bullet, residues of primer and propellant
chemicals from the cartridge are propelled both forwards
toward the target and backwards toward the firer. The
disposition and subsequent analysis of these residues can be
used to (a) help in identifying the firer, or persons close to the
firer, and (b) estimate the distance from which the weapon was
fired.
Primers tend to be inorganic, and propellants organic
compounds. A typical primer consists of lead styphnate
(explosive), antimony sulfide (fuel) and barium nitrate
(oxidizer). The propellant is usually a smokeless powder
containing nitrocellulose. There are different types of

This journal is ß The Royal Society of Chemistry 2005 Chem. Soc. Rev., 2005, 34, 1021–1030 | 1021
propellants, those termed ‘‘single base’’ which consist only of
nitrocellulose, and ‘‘double base’’, containing nitrocellulose
and nitroglycerine. There are triple base propellants, contain-
ing nitrocellulose, nitroglycerine and nitroguanidine, but they
are usually restricted to large calibre ammunition. Complete
combustion of the propellant is rare, and organic residues
consist of unburnt or partially burnt nitrocellulose and
nitroglycerine. Inorganic residues are formed from condensa-
tion of vapours from the hot primer compounds, and comprise
hollow spherical particles, whose elemental composition (Pb,
Sb, Ba) is critical to their identification as firearms discharge
residues.
Collection of the residues is made in a number of ways, e.g.
swabbing and vacuuming, and taping. The most convenient
method for collecting primer residues is by ‘‘taping’’ the hands
and forearms of suspects with adhesive tapes. Analysis4 of the
tapes is undertaken in a scanning electron microscope with
energy dispersive spectroscopy (SEM-EDS) for the presence of
lead, antimony and barium in individual particles. The
morphology of the particles as hollow spheres in combination
with their elemental composition identifies them uniquely as
firearms discharge residue particles. Lead-free ammunition
(Sintox) has zinc and titanium as its main elements, and after
discharge, the similar morphology of spheroidal particles
together with elemental composition Zn and Ti distinguishes
them from environmental particles of the same composition.
Swabbing and vacuuming is used for recovery of both
propellants and primers. Swabs and micro-filters are extracted
for propellants and retained for primer analysis. Analytical
methods4 for propellants, with detection limits of 100 pg to
1 ng, include high performance liquid chromatography
(HPLC) with amperometric detection, gas chromatography
with thermal energy analysis (GC-TEA), and liquid chroma-
tography with mass spectrometry (LC-MS).5 A recent method6
for dual analysis (organic and inorganic components) has been
suggested using adhesive tape.
Distinguishing between genuine firearms discharge residue
particles and those of aggregates of environmental occupa-
tional origin is important. The possibility of occurrence of
other, non-firearms residue particles with elemental content
Pb, Sb and Ba has encouraged research and various surveys.
Of particular interest is the elemental composition of
automobile brake pads7, pyrotechnics7,8 and fireworks.
Another aspect of FDR analysis concerns detection and
identification at the crime scene. To this end, portable X-ray
fluorescence (XRF) equipment9 for general crime scene work
has been developed and successful detection of FDRs and
identification of Sb, Pb and Ba has been achieved on a variety
of surfaces.
Toxicology
The forensic toxicologist’s role2,3 is to detect and identify the
presence of drugs and poisons in body fluids, tissues and
organs. As in the detection and analysis of firearms discharge
residues, the amount of material the toxicologist is expected to
work with is low, being of the order of nanograms or at best
micrograms. Widespread use and abuse of drugs means that
the majority of toxic materials are drugs, so the analyst has to
1022 | Chem. Soc. Rev., 2005, 34, 1021–1030
devise a means of extracting the drugs and their metabolites,
identifying them, and if necessary quantifying them.
Occasionally, the forensic toxicologist has to deal with cases
of heavy metal poisoning. The metals may have to be extracted
from body fluids or from contaminated foodstuffs, and
identified by techniques such as X-ray fluorescence.10
When the type and/or number of drugs in the toxicological
sample is unknown, a screening procedure, for example
enzyme immunoassay (EIA), is initially employed. Screens
for opiates, cannabis, amphetamines, cocaine, methadone and
benzodiazepines are usually run as a matter of routine,
together with any others if prior knowledge indicates specific
drug usage. Positive screening results provide the analyst with
a list of drugs to be extracted, confirmed and quantified. A
fresh sample is taken and the drugs of interest are then
extracted.
Drugs are classified broadly as acidic (e.g. barbiturates),
basic (e.g. the amphetamines, cocaine and methadone), and
neutral or amphoteric (e.g. morphine and the opiates). They,
and their main metabolites, are extracted from body fluids by
controlling the pH of the aqueous solution in which they are
dissolved. Solid phase extraction (SPE) is widely used now in
preference to liquid/liquid extraction, mainly because it is
amenable to automation, and SPE equipment manufacturers
will often provide method sheets for users. SPE cartridges may
be used for small scale bench-top work, or for larger scale
automated process operation. After extraction and clean-up,
the presence of the drug(s) is confirmed by a spectroscopic
method, either Gas Chromatography-Mass Spectrometry
(GC-MS) or LC-MS. Confirmation by either of these
methods11 is essential, because the screening technique may
not be specific enough. These hyphenated techniques are
highly specific and they enable quantification to the
required levels of sensitivity. GC-MS methods generally
require derivatisation of the extracts in order to render them
volatile, whilst LC-MS methods do not. LC-MS is
particularly useful when the compounds in question are
thermally labile.
Driving whilst under the influence of drugs is a problem, the
seriousness of which has only relatively recently been
recognized. In the UK between 1985 and 2000, the proportion
of road traffic fatalities with illicit drugs in their bodies12
increased from 3% to 18%. This research evidence supported a
growing public perception that ‘‘drug driving’’ was contribut-
ing to death and injury on the road. The Police Service has
adapted standardized field sobriety tests (SFST) and drug
recognition examinations (DRE)3 for use in the UK, and Field
Impairment Testing (FIT) was introduced in 2000. In 2001 the
UK Forensic Science Service (FSS) introduced a streamlined
analytical procedure13 which enabled it to process a large
number of motorists’ samples following positive FIT. This is
based upon the procedures described above: screening,
extraction and confirmation. The use of oral fluid, as opposed
to blood or urine, for the detection of drugs of abuse has
increased. The collection of oral fluid is non-invasive and the
testing detects primarily the parent drug and lipophilic
metabolites. It has the potential to provide a convenient,
rapid, roadside detection system14 similar to that used to detect
alcohol. A minimum of 1 ml oral fluid enables roadside
This journal is ß The Royal Society of Chemistry 2005
immunoassay screening with sufficient fluid left for laboratory
GC-MS, LC-MS, or LC-MS/MS confirmation.
Drug-facilitated sexual assault, or the use of so-called ‘‘date
rape’’ drugs, is a crime in which the perpetrator surreptitiously
administers drugs (usually through alcoholic drinks) to the
victim prior to a sexual assault. The drugs work in a way that
renders the victim physically helpless and unable to remember
what has happened. There are three well-known date rape
drugs: GHB (gamma hydroxybutyrate), Rohypnol (flunitra-
zepam) and Ketamine (ketamine hydrochloride). Rohypnol is
legal in Europe and is prescribed for sleep problems and as an
anaesthetic, but it is illegal in the United States of America.
Ketamine is an anaesthetic for humans and for animals, and
GHB is also used to treat sleep problems. Other hypnotic
drugs such as Zolpidem15 and Zopiclone16 have also been
reported in cases of date rape. The low levels of these
substances have been detected and measured using LC-MS/
MS. Despite all the publicity given to date rape drugs, in a
substantial proportion17 of alleged date rape cases it was found
that the only drug involved was alcohol.
Drug analysis
The main types of drugs of abuse2,18 likely to be encountered
by a forensic drugs analyst are: cannabis, amphetamines,
benzodiazepines, heroin, and cocaine. They appear in sub-
gram quantities as so-called ‘‘street seizures’’ in the possession
of individual users, in larger amounts in the hands of local
drug dealers, and in kilogram quantities as imported drugs
(mainly cannabis, heroin and cocaine). With regard to their
analysis, the forensic scientist’s main tasks are to (a) determine
whether or not a controlled substance is present, (b) determine
how much of the substance is present, and (c) determine, on
occasion, the relationship of drug samples to each other19
through comparison or ‘‘profiling’’.
Cannabis samples submitted to the laboratory can be in one
of three forms: herbal material, resin and oil. The main
physiologically active ingredient of cannabis is D9-tetrahydro-
cannabinol (D9-THC). Identification of herbal cannabis is
achieved by visual inspection under low power magnification.
Ethanol extracts of the herbal material, resin and oil are
subjected to thin layer chromatography (TLC) for rapid
screening and simple comparison purposes. For identification
and confirmation, trimethylsilyl derivatives of the main
components D9-THC, cannabidiol, cannabinol and the lesser
components D8-THC and D9-tetrahydrocannabinolic acid are
analysed by GC-MS. The technique will identify, unequi-
vocally, the derivatised compounds. HPLC or GC-MS can be
used for profiling purposes. The preliminary screen by TLC
will provide a good indication as to whether or not the blocks
of resin are from the same batch, and reversed-phase HPLC
will confirm this. HPLC is particularly useful because unlike
GC-MS, it does not require the samples to be derivatised.
Tetrahydrocannabinolic acids are thermally labile and would
decompose under GC-MS conditions. The chromatogram
serves as the profile of the drug.
The title ‘‘amphetamines’’ includes amphetamine itself,
methylamphetamine, 3,4-methylenedioxyamphetamine (MDA),
3,4-methylenedioxymethylamphetamine (MDMA, Ecstasy) and
This journal is ß The Royal Society of Chemistry 2005
3,4-methylenedioxyethylamphetamine (MDEA). These drugs
are usually synthesized, and when sold as powders are
adulterated or diluted with a variety of compounds
including glucose monohydrate, mannitol, Epsom Salts
(MgSO4?7H2O), caffeine and starch. The drugs are usually
taken orally, in either powder or tablet form. The Marquis
test19 is a useful presumptive test for amphetamines and
other drugs, relying on a colour reaction. In the presence of
amphetamine and methylamphetamine, a yellow-orange
coloration results. With ring-substituted amphetamines, a
blue-purple coloration is obtained. TLC is then used to
determine the amphetamine present.
Confirmation of the drugs is by GC-MS, with suitable
derivatisation. Nuclear magnetic resonance (NMR)20 or
HPLC is used to quantify the drugs, but comparison of
samples requires GC-MS. The methods of synthesis of the
drugs are well known, and levels of impurities expected for
each route are known. These impurities which constitute the
‘‘fingerprint’’ or profile are numerous and not all have been
identified, but the resulting chromatograms from samples are
easily compared visually, and the components identified by
the mass detector. Isotopic characterization of MDMA using
the d15N ratio successfully discriminated between batches of
the illicit material. Raman spectroscopy21 has also been used
for composition profiling of Ecstasy tablets. Far-red excitation
(785 nm) has been investigated as a simple and rapid
technique for MDMA tablet profiling. The spectra obtained
are rich in vibrational bands and permit identification of the
active drug and excipients used to bulk the tablets. Relative
band heights can be used to determine drug/excipients ratios
and the degree of hydration of the drug. Some 50 tablets per
hour were analysed, demonstrating the high throughput
nature of the technique. In one case, a sample of 400 tablets
from a seizure of .50,000 could be classified on the basis of
excipients used. Some were diluted with sorbitol, some with
cellulose and the remainder with glucose. Further, more
detailed analysis revealed differing drug/excipient ratios
within a particular drug/excipient combination, and also a
difference in the degree of hydration in the MDMA feedstocks
used to manufacture the cellulose-, glucose-, and sorbitol-
based tablets.
Benzodiazepines, unlike heroin, cocaine, cannabis and
amphetamines, are controlled pharmaceutical drugs. They
are usually encountered as tablets or capsules containing
controlled amounts of drug. A seizure may consist of a few, or
hundreds or thousands of capsules, and a representative
number19 will be selected for analysis. A detailed description
of the capsules or tablets is made first, and compared with
manufacturers’ information, and then analysis is undertaken.
Presumptive tests, if positive for benzodiazepines, are followed
by thin layer chromatographic analysis.
Benzodiazepines do not require derivatisation and are
straightforward to analyse by GC-MS. If chromatographic
retention times and spectral information match, then the
identification is confirmed. Quantification is conveniently
obtained through HPLC with UV detection. Profiling of such
compounds is not appropriate because the drugs are manu-
factured under strict quality control conditions, and will be of
high purity.
Chem. Soc. Rev., 2005, 34, 1021–1030 | 1023
 Heroin, or diamorphine (diacetylmorphine), is produced by
acetylating morphine which is isolated from the opium poppy.
The occasional ‘‘vinegary’’ smell of a powder containing
heroin is due to hydrolysis and the subsequent formation of
monoacetylmorphine, morphine and acetic acid. The purity of
street level drugs varies considerably, but the average street
seizure contains 35 wt% diamorphine. The most common
forms of heroin are the salt (the hydrochloride hydrate), which
is water soluble, and the base which is insoluble in water. The
salt is therefore the form of choice for injection, whilst the base
is usually snorted or smoked. A decade’s worth (1980–89) of
heroin analysis22 by X-ray powder diffraction (XRD) revealed
an interesting trend, in that street seizures containing the
hydrochloride were prevalent at the beginning of the decade,
but its use was supplanted by that of the base by the end of the
period. The decade concerned also happened to be that in
which awareness of the disease AIDS was raised, so one may
speculate that the move by addicts from injection to smoking
the drug was motivated in part by a desire to avoid contracting
the disease through use of shared hypodermic needles.
Street seizures comprise the drug, as salt or base, mixed or
‘‘cut’’ with various materials such as phenobarbitone, para-
cetamol, caffeine, procaine, lignocaine, codeine, phenolphtha-
lein, calcite and lactose monohydrate. Heroin itself usually
consists of diamorphine, 6-o-monoacetylmorphine, morphine
and codeine, so most ‘‘heroin’’ samples are quite complex if
adulterated with some of the aforementioned compounds. The
usual presumptive test for heroin is Marquis Reagent, with
which opiates react19 to give a blue-violet colour. Thin layer
chromatography with a suitable elution solvent reveals most of
the components of the mixture, whilst GC-MS is the method
of choice again for heroin confirmation/identification.
Quantification of heroin samples may be achieved by either
GC or HPLC. GC of course will require derivatisation of
samples, but HPLC requires only the dissolved samples. Either
method proceeds with the simultaneous analysis of sample
complete with internal standard, and then analysis of a set of
calibration standards whose concentration range encompasses
the range of heroin concentrations anticipated. If comparison
of heroin seizures is required, the comparison has to be both
qualitative and quantitative, and GC-MS is the method used.
TLC will suffice as a simple means of comparison, and it is
most effective for a large number of samples in that it will
quickly identify samples that are not similar, but it is not
quantitative. HPLC is quantitative but does not provide the
analyst with a definitive identification of each chromato-
graphic peak. Comparison of samples demands exactly the
same treatment of each sample, and if, as a result, the
chromatograms and mass spectral data are indistinguishable,
the samples are deemed to have come from the same source.
Cocaine is listed as a Class A drug in schedule 2 of the
Misuse of Drugs Act 1971.18 It occurs naturally in plant
material and is extracted from coca leaves. One form of
cocaine that has gained widespread popularity is known as
crack. It is the free base form of cocaine and is prepared from
cocaine hydrochloride by mixing the salt with sodium
bicarbonate and water, and heating it. Crack is smoked or
snorted. The salt, cocaine hydrochloride is a white powder
known as snow. Both the hydrochloride and the base are sold
1024 | Chem. Soc. Rev., 2005, 34, 1021–1030
on the street as powders cut with various adulterants such as
mannitol, glucose monohydrate, and local anaesthetics ligno-
caine and procaine base and hydrochloride. Presumptive
tests19 for cocaine include the cobalt isothiocyanate test, which
is not completely cocaine specific. This test, and a modified
version, called the Scott test, which is more specific, are both
based on reactions which produce colour changes. TLC is
performed on methanol solutions of the street seizures, and
will identify cocaine, its related compounds and lignocaine and
procaine. GC-MS is used to confirm the presence of cocaine
and also to quantify it.
Cocaine, like heroin, is produced in batches whose
impurities, and level of impurities, will differ. As a result,
seizures from different batches are unlikely to have the same
overall composition at trace level, and the makeup of a sample
can act as a ‘‘signature’’ for its batch, and in some cases,
indicate its geographical source. Cocaine profiling, like heroin
profiling, can result in the linking of seizures and identification
of probable source. The cocaine ‘‘profile’’ includes cocaine,
ecgonine, benzoylecgonine, nor-cocaine, the truxillines and
cinnamoyl cocaines. An account of the recent advances in
cocaine profiling methodology23 provides an interesting insight
to the subject.
Stable isotope ratio mass spectrometry (SIRMS) is an
analytical technique which has found wide application11,24 in
forensic science. It is appropriate to mention it in some detail
here, because its use in drug analysis is of considerable
importance. The sample to be analysed has first to be
converted into simple gases e.g. CO2, N2, CO, H2 and SO2,
and these gases are led to a dual input mass spectrometer
together with gas produced from a reference material. Gas
isotope ratio mass spectrometers are single focusing magnetic
sector instruments in which the isotopes are detected
continuously and simultaneously by a multi-collector array.
Stable isotope measurements are expressed in delta (d) values
according to the formula:
d13C(%) 5 ((R13sample 2 R13standard) 6 103)/R13standard
where R13 5 13C/12C.
The abundance of the largest trace isotope of carbon (13C) is
11,000 ppm (0.011) relative to the major isotope (12C). Other
abundances are: 2H/1H 5 158 ppm, 15N/14N 5 3,700 ppm,
18
O/16O 5 2,000 ppm and 34S/32S 5 42,000 ppm. Positive delta
values indicate that there is a greater percentage of the heavier
isotope present relative to the standard. Cocaine originating
from different geographic regions in South America25 can be
identified by its isotope ratio signature. By combining the
carbon (d13C) and nitrogen (d15N) isotope ratios with
detectable differences in the patterns of trace alkaloids, the
source of cocaine samples can be correctly identified. A similar
approach has been adopted for sourcing heroin.
Explosives
The majority of chemical explosives (energetic materials)26,27
contain oxygen, nitrogen and oxidizable elements (fuels) such
as carbon, hydrogen, and sulfur. Explosives may be classified
in terms of their chemical composition (those containing
molecular groups such as peroxides, ozonides, nitrates, nitrites,
This journal is ß The Royal Society of Chemistry 2005
chlorates, perchlorates, azides, fulminates and acetylides), but
a more useful classification is by their performance and uses.
Three convenient classes are: primary explosives, secondary
explosives and propellants.
Primary explosives differ from secondary explosives in that
they undergo a very rapid transition from burning to detonation,
and they have the ability to transmit the detonation to less
sensitive explosives. On detonation, the primary explosives
produce a great deal of heat and shock, which can be used to
initiate a secondary, more stable explosive. For this reason, the
primary explosives are used as initiators, or detonators. Primary
explosives are sensitive to detonation through shock, friction,
electric spark or high temperatures and will explode whether
they are confined or unconfined. Examples of primary explosives
are: lead styphnate (lead trinitroresorcinate), lead azide, tetra-
zene, and mercury fulminate.
Secondary explosives (high explosives) are less sensitive than
primary explosives and are only detonated by the shock
produced by the explosion of a primary explosive. A secondary
explosive, on detonation, decomposes instantaneously into
other more stable compounds (usually gases). RDX decom-
poses into carbon monoxide, steam and nitrogen. Examples
of secondary explosives are: nitrocellulose, TNT (trinitroto-
luene), nitroguanidine, picric acid, tetryl (trinitrophenyl-
methylnitramine), RDX (cyclotrimethylenetrinitramine),
PETN (pentaerythritol tetranitrate), HMX (cyclotetramethy-
lenetetranitramine), and TATB (triamino-2,4,6-trinitroben-
zene). Propellants are combustible materials containing
within themselves all the oxygen needed for their combustion.
They can be initiated by a flame or a spark, and change from a
solid to a gaseous state relatively slowly, i.e. in milliseconds.
Black powder (potassium nitrate, charcoal and sulfur) and
smokeless powder (nitrocellulose) are examples of propellants.
A forensic chemist can be faced with a number of
requests2,28 with regard to explosives. He/she may be asked
to identify (a) bulk material, as in the case of seizure of an
unexploded device, or (b) traces of explosives in order to
establish links between suspect(s) and the crime scene. Such
traces can occur in almost any setting—open spaces, commer-
cial buildings and private dwelling places, in and on motor
vehicles, on public transport systems, on individuals, their
clothing, and their private property. Analysis of post-blast
residues is also important, enabling the scientist to identify the
explosives that caused the blast, together with their likely
origin. Systematic procedures3 are followed in order to recover
whatever evidence may be present, both in linking suspect and
scene, and following an explosion.
Recovery of traces of explosives, or explosives residues, is
achieved by swabbing (dry or with a solvent), solvent washing
of items, vacuum sampling, or by adhesive tape. A rapid and
convenient screening method at the crime scene or in the
laboratory is ion mobility spectrometry29 (IMS). A portable
IMS instrument uses a vacuum to collect explosive residues
from suspect surfaces. Alternatively, the surface suspected of
containing explosive residues is wiped down with a cellulose or
Teflon filter disc, and the collected residues are then sucked
into the spectrometer. The residues are vaporized by the
application of heat, and converted into electrically charged
molecules or ions by means of metal foil containing 63Ni,
This journal is ß The Royal Society of Chemistry 2005
whose b-particles initiate ionization of the air. The ions then
pass along a cylindrical drift tube under the influence of an
electric field of y200 V cm21. The ions traverse the drift
region in a few milliseconds, and the detector signal from the
different ions with characteristic drift times constitutes the
mobility spectrum.
IMS permits fast, highly sensitive and specific detection, but
relies upon the measurement of mobility as the sole
characteristic for identification, so the results of an IMS
screening test need to be confirmed. Residues submitted to the
laboratory are first examined microscopically and then
dissolved in a solvent such as ethanol or acetone prior to
colour spot tests and chromatographic analysis. The combina-
tion of the separation power of HPLC with the identification
capability of mass spectrometry (MS) is the most widely used11
analytical system. The high explosives TNT, NG, PETN and
RDX are efficiently ionized under negative ion atmospheric
pressure chemical ionisation (APCI) conditions.30 The limit of
detection is improved, in some cases by several orders of
magnitude, by complexation with chlorine, demonstrating this
to be a highly suitable method for enhancing the detection
capabilities for explosives.
Any detection of explosives or explosive residues must take
into account the background level at a specific site. A recent
survey31 of background levels of explosives in public places
was undertaken in four major UK cities: Birmingham, Cardiff,
Glasgow and Manchester. The survey concentrated mainly on
public means of transport (taxis, buses and trains), but also
included samples collected from airports, hotel rooms, private
houses, private vehicles, and clothing purchased from charity
shops. Samples were collected by wiping non-porous surfaces
with swabs impregnated with 1 : 1 ethanol : water, and by
vacuuming porous surfaces. GC-TEA and GC-MS were used
to detect, identify and confirm the presence of traces of NG,
TNT, PETN and RDX. LC-MS was used to analyse samples
for the presence of cyclotetramethylene tetranitramine (HMX).
Only one low-level trace (7.5 ng) of RDX was detected on a
collective swab of nine train seats. Nitroglycerine (3.6 ng),
possibly associated with the use of firearms, was found on the
floor of a taxi, and 2,4-dinitrotoluene (15.2 ng) was detected on
the back of an X-ray machine at Glasgow airport. The overall
conclusion was that it is unlikely that persons visiting public
areas could become significantly contaminated with explosives.
Classical trace evidence
This refers to evidence2,3 such as textile fibres, paint, glass and
soil. Prior to the advent of DNA profiling, these materials
constituted the main types of so-called trace evidence. The
search for evidence at a crime scene, or on a victim’s or
suspect’s clothing for textile fibres, paint flakes, and glass
fragments is highly labour-intensive, and some might say not
cost-effective. In the absence of DNA evidence and finger
marks, however, trace evidence of this nature may be the only
means of solving of a crime.
Textile fibres. Textile fibres may be classified in broad terms
as either natural or man-made. Further subdivision of
natural fibres leads to animal, vegetable and mineral fibres.
Chem. Soc. Rev., 2005, 34, 1021–1030 | 1025
Sub-classification of man-made fibres leads to: synthetic
polymer (polyester, polyamide, polyurethane, polyolefin,
polyvinyl), and natural polymer (rubber, cellulose ester,
regenerated cellulose, regenerated protein). Textiles are ubi-
quitous and they can be involved in crimes32 in many ways,
and can occur as evidence also in a variety of ways.
Forensic examination of any trace evidence, including textile
fibres, is based upon Locard’s Exchange Principle. This states
that ‘‘every contact leaves a trace’’. In practice, even though a
transfer of material has taken place (in one or both directions),
it may be impossible to detect, because the amount transferred
may be so minute. Also, some surfaces, because of their
disposition and texture, may shed transferred material easily
and quickly. As a consequence it is important to collect
clothing from suspects and victims as soon as possible after an
alleged offence, because evidence of contact (and hence
association) found through comparison of fibres, will generally
involve recent transfers.
Recovery or retrieval of fibres from a crime scene is achieved
through the use of forceps when they are easily visible, through
adhesive tape lifts, or vacuuming. The procedure for examina-
tion and analysis of fibres consists of microscopic comparison,
fibre identification and colour analysis. Known, or control
fibres are subjected to UV/visible comparison microscopy with
those extraneous (suspect) fibres recovered loose or from tape
lifts. The fibre type (man-made or natural) can be determined,
with morphological detail from optical microscopy usually
sufficient for identification of natural fibres, and Pyrolysis Gas
Chromatography (PGC) and Pyrolysis Mass Spectrometry
(PyMS) for identification of man-made fibres. If, after
comparison microscopy there are similarities, known and
suspect fibres are examined by visible light microspectropho-
tometry. If the resulting spectra are similar, FTIR examination
follows, which yields unequivocal fibre polymer identification
and some information concerning the dyes.
Dye extraction is the next stage, and the methods used will
depend upon the type of fibre and dye used. Solvent
extraction, enzymatic hydrolysis, and alkaline hydrolysis are
techniques used for the release of dyes from the various types
of fibre. Visually similar colours may consist of different
component dyes (a so-called ‘‘metameric match’’) which can be
readily distinguished by TLC. Thin layer chromatography
does have its limitations, particularly with pale yellow dyes and
with low concentrations of dye, and HPLC32 and Surface
Enhanced Resonance Raman Scattering Spectroscopy
(SERRS)32,33 have been used to good effect. SERRS33 is a
combination of Resonance Raman Spectroscopy (RR) and
Surface Enhanced Raman Scattering (SERS), and is a very
sensitive technique, possibly up to ten orders of magnitude
more sensitive than traditional Raman spectroscopy. Silver
colloids are used to obtain the surface enhanced effect.
Aggregation of these colloid particles and ensuring that the
analyte molecules are in close proximity to the colloid surface
are essential for achieving high sensitivity and reproducible
SERRS spectra.. An organic compound such as poly(L-lysine)
has proved to be an excellent aggregating agent. For fibre dye
analysis, a single strand of fibre is pre-soaked in sodium
hydroxide, rinsed and then soaked in an aqueous solution of
poly(L-lysine), rinsed again, and then treated with the
1026 | Chem. Soc. Rev., 2005, 34, 1021–1030
concentrated silver colloid. The fibre is placed on a microscope
slide and the SERRS spectrum of the colorant is recorded.
This in situ analysis of the dye is worthy of note because the
fibre remains intact (the method can be considered to be non-
destructive) and can be retained for further complementary
methods of examination. LC-MS has more recently been used
to identify dyes extracted from textile fibres.34
Just as with explosives, there is a need to take into account
the level of background fibre ‘‘contamination’’. The common
occurrence of textiles in daily life constitutes a major source of
background fibres. Knowledge of the frequency of occurrence
of fibre types in a given population is required in order to
assess the evidential value of finding fibres that could not be
differentiated from a suspect source. For example, studies of
the population of coloured fibres in human head hair,35
revealed that natural fibres (mainly cotton) at 72% were the
most prevalent with the balance of man-made fibres. Black/
grey cotton (48%), blue cotton (29%) and red cotton (y13%)
were the most common colours. A similar survey, but in a
different country, looking at the population of coloured textile
fibres in domestic washing machines36 revealed strikingly
similar results in that cotton fibres (y70%) were the most
prevalent followed by man-made fibres (y24%). Black/grey
cotton (27%), blue cotton (20%) and red cotton (y16%) were
the most common colours.
In general, the more common the fibres, the lesser the value
of the evidence. Conversely, fibres which are rare by virtue of
an unusual morphological characteristic, specific usage,
limited production or obsolescence will have strong evidential
value, even when they are present in low numbers.
Paint. Paint37 has a dual role, and that is to decorate and to
protect. The protective role is to shield the surface from its
environment. Decorative household paints must have proper-
ties that protect against the sun (ultraviolet radiation), rain
(moisture) and the atmosphere (oxygen), whilst vehicle paints,
as well as being required to resist the elements, must resist
chemical attack from salt, fuel and lubricants. It is clear that
these two types of paints, together with tool paints and
specialist paints such as security and anti-climb paints, will
therefore have differing compositions. Irrespective of minor
differences, however, all paints are composed of pigments,
extenders and binders as well as other additives, dissolved or
dispersed in a solvent. After the paint has been applied to a
surface, the solvent evaporates, leaving behind a hard
polymeric binder and any pigments that were suspended in it.
Paint occurs as trace evidence38 in a number of circum-
stances. It may occur as small flakes on the outer garments of a
person who has broken into property and damaged paintwork,
or as smears of vehicle paint transferred from one vehicle to
another in a traffic accident, or from a vehicle to a victim in a
hit-and-run accident. If a tool such as a jemmy has been used
in a break-in, it may well have smears of paint on its tip. In
some cases, usually hit-and-run cases, enough information
may be obtained from a paint flake left on the victim, to enable
identification of the colour, make, and model of the vehicle
involved. This does require cooperation from the vehicle
manufacturer, who will have records of paint batch composi-
tions and layer sequences.
This journal is ß The Royal Society of Chemistry 2005
 The analytical approach to such evidence begins with
microscopic examination of control paint, for example from
the point of entry in the case of a break-in, and suspect samples
(found on the garments of the suspect). Polarized light
microscopy and fluorescence microscopy are used as well as
visible light microscopy because much information can be
obtained concerning the overall appearance of the samples—
the presence of a layer structure, the colour and texture of
the layers, particle size information, and the presence of
contamination. Most forensic paint specimens encountered are
single layer, so after colour comparison, the chemical
composition of the pigments, extenders and binders must be
determined.
Colour comparison is usually undertaken by microspectro-
photometry. It is important that the control paint specimen be
taken from as close as possible to the suspect sample, to
eliminate differences in top-surface measurements caused by
light, weathering and ageing. Pigments can be organic or
inorganic, but only inorganic pigments are used for black or
white paints. Extenders, like pigments are particulate in
nature, and are usually cheap, synthetic inorganic materials
or minerals, and they are present to add bulk to the paint.
Analysis of paint pigments and extenders is undertaken by
Fourier transform infrared spectroscopy (FTIR), X-ray
powder diffraction (XRD), X-ray fluorescence (XRF) and
Scanning electron microscopy/energy dispersive spectrometry
(SEM/EDS). Scanning electron microscopy is particularly
useful because it provides a magnified image of the paint
flake specimen, together with the capability of elemental
analysis. The electron beam can be focused directly on
individual paint layers and on individual particles, and an
elemental analysis obtained. Inferences can be drawn as to
which extenders or pigments are present in the specimen, but
SEM/EDS alone will not provide definitive pigment identifica-
tion. XRD is a useful non-destructive method22 for paint
analysis in that it does provide definitive identification of the
pigments (organic or inorganic) and extenders, although
multilayer paints require separation of the layers prior to
analysis. Its chief drawback by comparison to the other
techniques is its relative insensitivity. Laser ablation-
inductively coupled plasma-mass spectrometry (LA-ICP-MS)
has also been used39 for elemental analysis. The laser ablation
method is capable of simultaneously sampling several layers
directly before analysis by ICP-MS for the identification and
quantification of trace elements present in different layers of
the sample. Trace element analysis adds a further means of
discrimination. Analysis of the binders in a paint specimen is
undertaken by pyrolysis methods38 such as Pyrolysis Gas
Chromatography (PGC) and Pyrolysis Mass Spectrometry
(PyMS), and FTIR.
Glass. Glass that is broken during the commission of a
crime38 can be a useful source of trace evidence, linking a
suspect with the crime scene. Splinters or fragments of glass
may be found in a suspect’s hair, on his outer garments, or
embedded in his shoes as a result of breaking and entering. In a
hit-and-run accident, fragments of glass from a broken
headlamp, like paint fragments, found on the victim, may
help to identify the vehicle involved.
This journal is ß The Royal Society of Chemistry 2005
Glass is an amorphous material, consisting of silicon oxides
mixed with metal oxides such as sodium, calcium, magnesium
and aluminium. The combination of these oxides is used in the
manufacture of the most common types of glass—window and
bottle glass. The forensic scientist will generally classify glasses
as sheet (window or ‘‘float’’), container (bottle, jar), tableware
(including lead glass), vehicle window, and vehicle headlamp
(borosilicate glass).
The majority of glass evidence usually consists of small
fragments recovered from clothing, and the task of comparing
them with a control sample from a window or bottle, for
example, comes down to physical and chemical analysis. The
physical properties of density and refractive index (RI) have
been used for characterizing glass fragments for many years,
whilst SEM-EDS, XRF and inductively coupled plasma
atomic emission spectroscopy (ICP-AES) and LA-ICP-MS
are used for elemental analysis. Refractive index is measured
by an automated system3,38 called GRIM (glass refractive
index measurement).
Elemental analysis38 is the final stage in glass comparison
and this will enable discrimination of glasses similar in density
and refractive index. Glass fragments are mounted in resin
blocks, polished, and examined in the SEM where energy
dispersive spectrometry yields elemental concentrations. XRF,
and in particular micro-XRF which employs capillary optics to
produce an intense, focused beam of X-rays, is used on small
irregularly-shaped fragments of glass. This experimental con-
figuration enables quantitative elemental analysis normally
associated only with flat polished surfaces of the fragments in
the SEM. Total reflection X-ray fluorescence (TXRF) is another
variant of conventional XRF, which uses grazing incidence
(instead of the usual 45u) for the incoming X-ray beam. The
signal to noise ratio is improved and dissolved glass samples
yield quantitative results as good as any other technique.
Of the two inductively coupled plasma techniques, ICP-MS
is more sensitive than ICP-AES, and when combined with laser
ablation (LA-ICP-MS), is non-destructive. Samples for ICP-
AES have to be dissolved prior to analysis. Typical analyses of
float glasses and container glasses reveal similar amounts of Si
(y34%) and Na (y10%), but quite different levels of Mg
(y2.5% & 0.5% respectively) and Al (y0.4% & y1%
respectively). The detection limit of ICP-MS (0.01 mg l21) is
an order of magnitude lower than that of ICP-AES.
Soil. Soil2,3 has long been regarded as potentially one of the
most useful evidence types, but also as one whose significance
has been one of the most difficult to interpret. Soil or mud
sticking to a suspect’s shoes and clothes, and soil on a vehicle’s
tyres may be used to link a person or vehicle to a crime scene.
Forensic soil analysis, just as paint and glass analysis, is
comparative, with soil found on a suspect being compared
with that from a crime scene. Soil found on a suspect may
suggest a particular locality, because of its unique or unusual
chemical composition. This is specialist work and would
normally be undertaken by a geologist.
Procedures for soil examination will depend upon just how
much sample is available. Side-by-side comparison of control
and suspect samples (if they are bulk samples) will quickly
establish similarity or dissimilarity. Before colour comparison
Chem. Soc. Rev., 2005, 34, 1021–1030 | 1027
can be made, however, both samples must be dried thoroughly
because wet soils appear darker than dry soils. Low power
microscopic examination will reveal vegetation content, animal
remains and man-made debris. Greater magnification reveals
the mineral content of the soil. If only smears of soil make up
the suspect sample, then high magnification microscopy will
probably be the main means of comparison.
The real difficulty with soil as evidence is its compositional
variability. If the soil composition does not vary significantly
for a considerable area around the crime scene, then its
evidential value in linking soil on a suspect to soil at the scene
is very low. Even when variations in composition do occur
within metres of the scene, the significance of a match between
suspect soil and crime scene soil must be viewed with caution.
This is where detailed mineralogical analysis and trace element
analysis becomes important.40 Combining ICP-AES and ICP-
MS allows the abundance of up to 50 elements to be
determined in small samples. For most elements, ICP
measurement precision is good, but a degree of variation can
arise as a result of sub-sampling procedures, and selective
transfer mechanisms relating to forensic samples as well as the
degree of spatial variation existing in nature. If the sample size
is sufficiently large, the analysis of several different size
fractions can be attempted, but usually analysis of a
standardized ,150 mm fraction separated from a bulk sample
provides sufficient discrimination between samples, and is the
most practical means for the mass screening of samples. Stable
isotope ratio variation in soils has been examined in order to
determine its usefulness in forensic soil investigations. Carbon
and nitrogen abundance and 13C and 15N isotopic values were
determined from six different locations using continuous flow
isotopic ratio mass spectrometry.
Other trace materials. In cases of assault, trace evidence such
as smears of cosmetics (face powder, lipstick, mascara, eye
liner etc.) and smears of shoe polish are occasionally
encountered. These materials usually contain wax-like sub-
stances together with pigments or dyes. The composition of
these materials can be obtained by methods including XRD,
XRF, SEM/EDS and SERRS. The in situ capability of
SERRS33 permits analysis of, for example, lipstick and shoe
polish smears without removing them from a substrate. An
aqueous solution of poly(L-lysine) is applied to the smear
followed by concentrated silver colloid solution which is
allowed to dry naturally. The SERRS spectrum is obtained by
excitation of the sample at 514.5 nm.
Arson
Fire investigation consists of establishing how, where, and
when a fire started. In reality arson is one of the most difficult
crimes to investigate.2,3 There is always the chance that the fire
could have been started accidentally, by a discarded cigarette
butt, or by a faulty electrical appliance, but the physical
evidence of this may well have been destroyed in the fire. Even
if it had been started deliberately, the link between a suspect
and the crime scene may be difficult to prove, and often the
consequences of extinguishing the fire may hamper the forensic
scientist’s investigation.
1028 | Chem. Soc. Rev., 2005, 34, 1021–1030
If the cause of the fire is thought to have been accelerants
(ignitable liquids), detection and identification of these
substances, or their residues, is one of the scientist’s first
tasks. Most arson attacks are started with accelerants such as
petrol (gasoline) or paraffin (kerosene), with occasional use of
paint thinners, and if any residues of these substances are left
after the fire has been extinguished, they may evaporate
quickly. It is important that the scientific investigation begins
as soon as possible. Since ignitable liquids always flow to the
lowest point, the most severe burning would be expected on
the floor rather than on the ceiling. The suspect may have
spread a trail of petrol or paraffin from a pile of debris to a
point of relative safety for himself, such that once the trail was
ignited, he could make good his escape without fear of injury.
Once the origin (seat) of the fire has been located, the
scientist may use a ‘‘sniffer’’ or portable vapour detector.
Accelerants are never totally consumed in the fire, because
they may be absorbed in flooring, carpets, plasterboard and
upholstery. The sniffer, whilst not specific in its identification,
detects combustible vapours, confirming the investigator’s
suspicions. Gas chromatography (GC) is the technique most
widely used to analyse traces of accelerants recovered from fire
scenes. The analysis starts with headspace sampling of the bags
or jars that contain the debris and clothing. The container is
heated, driving any volatile residue into its airspace. A syringe
containing an absorbent material, such as the resin Tenax, is
pushed through the container into the headspace air and it is
used to draw the air containing volatile compounds through
the resin. The absorbed volatiles are then thermally desorbed
from the Tenax when the sample tube is inserted in the GC.
The resulting chromatogram will comprise a mixture of peaks
from the accelerants and any pyrolysis products from the
thermal breakdown of plastics and natural materials due to the
heating effect of the fire. The more volatile components of
petrol will not be present in the residues recovered from a fire
scene, and at first this may hinder identification, but
comparison with chromatograms of standard petrol, paraffin,
diesel and white spirit will generally identify the accelerants.
If identification by GC alone proves difficult, GC-MS is
used in selective ion mode to identify the components of a
particular accelerant. Tandem mass spectrometry11 has also
been used for low levels of accelerants in the presence of a large
amount of background material. Gas chromatography/isotope
ratio mass spectrometry (GC-IRMS) technology41 has also
been used in an attempt to link the accelerant from fire debris
to accelerant identified in samples of the suspect’s clothing or
containers.
Inks
Forensic document examiners are mainly interested in the
identification of signatures and handwriting, but they are also
interested in writing materials and printing equipment. The
composition of inks, papers and the materials from which
documents are produced can be of great significance in a case.
With respect to the identification and interpretation of
alterations, deletions and additions to documents, a study of
the chemical composition of the ink2,3 used on documents may
confirm whether or not two documents were written using the
This journal is ß The Royal Society of Chemistry 2005
same pen. One way of comparing inks is by visible light
microspectrophotometry, and viewing the writing under
different conditions of illumination (infrared and ultraviolet
light) can also reveal differences in the inks. Thin layer
chromatography (TLC) is a simple and convenient way of
showing the multiple dye components normally present in
commercial ballpoint pen inks. As commercial ink formula-
tions tend to change over the years, it is possible to state with
some degree of confidence that certain documents are not as
old as they are claimed to be, if their inks are shown to contain
relatively modern chemical components.
Raman spectroscopy is used when inks cannot be distin-
guished using more traditional infrared and fluorescence
techniques. Surface Enhanced Resonance Raman Scattering
(SERRS)33,42 is used to obtain detailed spectra from ink
samples that are either weak Raman scatterers or show
dominant fluorescence when analysed by conventional
Raman spectroscopy. The in situ analytical capability is again
important here, as in textile fibre dye analysis, because the
technique is virtually non-destructive, but micro-application of
the colloid solution to the pen stroke under analysis is
essential.
Determination of the relative age of ballpoint pen inks,
written on the same paper with the same ink formulation, has
been and still is a controversial subject. Ink was extracted from
written paper samples taken from 1990 and 2000, made with
BIC blue and black pens, and the extracts were analysed
by LC-MS and LC-MS-MS methods5 in an attempt to
characterize the ageing of inks on paper. It has long been
known that 2-phenoxyethanol (PE), a volatile organic
compound found in ballpoint pen inks, evaporates as the inks
age. In a study of 633 ballpoint pen inks, GC-MS was used43 to
establish that PE occurs in 237 out of 279 (85%) black inks,
and in 293 out of 354 (83%) blue inks. In another study44 the
disappearance of PE was measured (with GC-MS) as a
function of time, and curves showing an exponential reduction
in PE were obtained for two ballpoint pens each with different
inks. Ink solvents were shown to evaporate rapidly, and
behave differently depending upon the type of paper used;
consequently the method was considered unsuitable for the
dating of ballpoint pen inks in forensic document examination.
Summary & future prospects
Forensic analytical chemistry has two main purposes, namely
identification and comparison. By-products of the identifica-
tion process are databases comprising analytical results,
qualitative and, where possible, quantitative. The importance
of analytical databases in forensic science cannot be empha-
sized enough, because they will influence the significance
placed upon a match between control and suspect samples. In
a world where mass production has largely taken over from
small, independent manufacturers of, for example, materials
such as paint and glass, stringent quality control means little
variation in chemical composition from batch to batch. As a
consequence, the search for, and identification of, new, more
sensitive chemical discriminators, has become important. Any
new analytical method offering greater sensitivity, however,
brings with it a concomitant increase in the dangers posed by
This journal is ß The Royal Society of Chemistry 2005
contamination, and method validation must take this into
account. Background information on the random occurrence
of textile fibres, glass, explosives and drugs in certain
environments forms another database that allows a level of
significance to be placed upon a particular analytical result.
Collection of this information can be tedious and time-
consuming, but it is important because it enables the forensic
scientist to have confidence in his/her assertion that the
presence of trace evidence, however small, is indeed real and
significant.
The future of any science is hard to predict, but there are a
number of areas in which forensic science will probably
advance, including miniaturisation,5 analysis at the crime
scene, and automation of laboratory analytical processes to
enable higher throughput of samples. Analysis at the crime
scene is currently undergoing trials45 in the UK with a mobile
laboratory for DNA analysis, footwear and fingerprint
information, and mobile phone data. Teleforensics, a terres-
trial version of the technology used by NASA for non-
destructive remote space (planetary) exploration capabilities,
has been applied in the United States to crime scene elemental
analysis9 for traces of firearms discharge residues, blood and
semen, using portable XRF equipment.
It has been said, not unkindly, that forensic science is not
‘‘rocket science’’, but the science behind forensic science most
certainly is, with the science behind DNA profiling and stable
isotope ratio mass spectrometry (SIRMS) as two shining
examples. Both techniques have their roots in fundamental
science, and their applications in forensic science lead, in their
own way, to specific identifications. Surface Enhanced
Resonance Raman Scattering Spectroscopy (SERRS), too,
shows considerable potential as a virtually non-destructive
method of in situ trace analysis. Just as the technique of DNA
profiling will surely develop further, SIRMS and SERRS
undoubtedly will, establishing themselves as key analytical
methods for determining, respectively, the source and compo-
sition of materials such as explosives, drugs, soils, glass,
pigments, dyes, inks, fuels, counterfeit money and many
others.
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